Rational engineering-based scalable production of Morus alba urease in Pichia pastoris optimized biofermenter for industrial silkworm rearing
Related Experiment Video
Updated: Oct 30, 2026

Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology
Published on: December 15, 2017
Marriam Khurshid1, Yufeng Zhao1, Justice Kwaku Addo2
1Jiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang, Jiangsu, 212100, PR China.
Abstract:
Developing a fermenter-compatible Morus alba urease with reproducible functional performance remains a challenge in sericulture industry. To address this challenge, M. alba urease gene (AB479106.1) was codon-optimized, subjected to rational design-guided protein engineering, and heterologously expressed in Pichia pastoris. Box-Behnken optimized parameters: media pH 6.5, 220 rpm agitation, and 0.75% (v/v) methanol, maximized extracellular urease activity. Purified synthetic wild-type (WT) enzyme exhibited a Kₘ of 1.60 mM, a kcat of ~10,141 s-1, and a specific activity of ~2967 U·mg-1, with high thermal stability (T50 = 71.4 °C) and optimal pH of 9, comparable to M. alba urease. Cytotoxicity assays on Bombyx mori ovarian cells and silkworm-associated microbiota confirm biocompatibility. Scale-up to 3 L fed-batch biofermenter significantly enhanced protein yield (2.4-2.7-fold), thermal stability (up to 1.4-fold), and productivity (~1.3-fold). This study presents an optimized P. pastoris biofermenter for sustainable production of biologically compatible plant-derived urease for scalable industrial sericulture applications.
More Related Videos
Related Concept Videos
Bioreactor Controls-III
Upstream Processing
Scale-Up Processes
Production of Alcohol
Production of Pharmaceuticals
Production of Antibiotics

